Chen Ping, Liu Xi-Qiao, Lin Xiang, Gao Li-Ying, Zhang Shuo, Huang Xuan
Department of Laboratory Medicine, The First Affiliated Hospital of Zhejiang Chinese Medical University, Hangzhou 310006, Zhejiang Province, China.
Department of Gastroenterology, The First Affiliated Hospital of Zhejiang Chinese Medical University, Hangzhou 310006, Zhejiang Province, China.
Mol Ther Oncolytics. 2021 Jan 16;20:228-239. doi: 10.1016/j.omto.2021.01.001. eCollection 2021 Mar 26.
Colorectal cancer (CRC) has a high mortality rate and poor prognosis. Despite chemotherapeutic agents such as cisplatin, which has achieved a better prognosis and survival rate against cancer, drug resistance leads to significant challenges. Accumulating evidence suggests that YTHDF1, the -methyladenosine (m6A) "reader," is an important regulator in tumor progresses. Herein, we report that YTHDF1 was significantly upregulated in human colon tumors and cell lines. Overexpression of YTHDF1 decreased the cisplatin sensitivity of colon cancer cells. From the established cisplatin-resistant CRC cell line (LoVo CDDP R), we detected that YTHDF1 was significantly upregulated in cisplatin-resistant CRC cells. Intriguingly, RNA sequencing (RNA-seq) results revealed that glutamine metabolism enzymes were clearly upregulated in LoVo CDDP R cells. Glutamine uptake, that is, glutaminase (GLS) activity, was upregulated in LoVo CDDP R cells. Furthermore, bioinformatics analysis indicated that the 3' UTR of GLS1 contained a putative binding motif of YTHDF1, and an interaction was further validated by a protein-RNA interaction assay (RNA immunoprecipitation [RIP]). Furthermore, we demonstrated that YTHDF1 promoted protein synthesis of GLS1. Inhibiting GLS1 effectively synergizes with cisplatin to induce colon cancer cell death. Finally, that YTHDF1 mediated cisplatin through the GLS1-glutamine metabolism axis was validated by an xenograft mouse model. In summary, our study reveals a new mechanism for YTHDF1-promoted cisplatin resistance, contributing to overcoming chemoresistant colon cancers.
结直肠癌（CRC）死亡率高且预后较差。尽管顺铂等化疗药物在对抗癌症方面取得了较好的预后和生存率，但耐药性带来了重大挑战。越来越多的证据表明，YTHDF1，即N6 - 甲基腺苷（m6A）“读取器”，是肿瘤进展中的重要调节因子。在此，我们报告YTHDF1在人类结肠肿瘤和细胞系中显著上调。YTHDF1的过表达降低了结肠癌细胞对顺铂的敏感性。从建立的顺铂耐药CRC细胞系（LoVo CDDP R）中，我们检测到YTHDF1在顺铂耐药CRC细胞中显著上调。有趣的是，RNA测序（RNA-seq）结果显示，谷氨酰胺代谢酶在LoVo CDDP R细胞中明显上调。LoVo CDDP R细胞中的谷氨酰胺摄取，即谷氨酰胺酶（GLS）活性上调。此外，生物信息学分析表明，GLS1的3'UTR包含一个假定的YTHDF1结合基序，并且通过蛋白质 - RNA相互作用测定（RNA免疫沉淀[RIP]）进一步验证了这种相互作用。此外，我们证明YTHDF1促进了GLS1的蛋白质合成。抑制GLS1可有效与顺铂协同诱导结肠癌细胞死亡。最后，通过异种移植小鼠模型验证了YTHDF1通过GLS1 - 谷氨酰胺代谢轴介导顺铂耐药。总之，我们的研究揭示了YTHDF1促进顺铂耐药的新机制，有助于克服化疗耐药的结肠癌。
